Shi Min, Zhao Shulin, Huang Yong, Zhao Limin, Liu Yi-Ming
Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education), College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin 541004, China.
Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education), College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin 541004, China.
Talanta. 2014 Jun;124:14-20. doi: 10.1016/j.talanta.2014.02.032. Epub 2014 Feb 28.
A signal amplification strategy based on antibody-gold nanoparticle-DNAzyme assembly in capillary electrophoresis based chemiluminescent immunoassays (CE-CLIA) was developed. In this CE-CLIA, antibody-AuNP-G-quadruplex/hemin was incubated with limited amount of antigen, and the formed immunocomplex and unreacted antibody-AuNP-G-quadruplex/hemin were then separated by CE and detected by CL. Due to the strong CL catalytic ability of G-quadruplex/hemin DNAzyme and a high loading ratio of DNAzyme on each AuNP, the assay was very sensitive. By taking carbohydrate antigen 19-9 (CA19-9), one of the most important carbohydrate tumor marker as the model analyte, the proposed CE-CLIA method for CA19-9 detection showed a linear range from 0.025 to 1.00 U/mL with a detection limit of 0.016 U/mL (signal/noise=3), which was more sensitive than the methods previously reported for CA19-9 quantification. The method was applied to quantify CA19-9 in human serum samples, and analytical results were in a good agreement with those obtained by using an established ELISA method.
基于抗体-金纳米颗粒-脱氧核酶组装的信号放大策略被应用于毛细管电泳化学发光免疫分析（CE-CLIA）中。在这种CE-CLIA中，抗体-金纳米颗粒-G-四链体/血红素与限量的抗原孵育，然后通过毛细管电泳分离形成的免疫复合物和未反应的抗体-金纳米颗粒-G-四链体/血红素，并通过化学发光进行检测。由于G-四链体/血红素脱氧核酶具有很强的化学发光催化能力，且每个金纳米颗粒上脱氧核酶的负载率很高，该检测方法非常灵敏。以最重要的糖类肿瘤标志物之一——糖类抗原19-9（CA19-9）作为模型分析物，所提出的用于检测CA19-9的CE-CLIA方法线性范围为0.025至1.00 U/mL，检测限为0.016 U/mL（信噪比=3），比先前报道的用于CA19-9定量的方法更灵敏。该方法被应用于定量检测人血清样品中的CA19-9，分析结果与使用既定酶联免疫吸附测定法获得的结果高度一致。
